Karl K. Haase

Prognostic Value of Plasma N-Terminal Pro-Brain Natriuretic Peptide in Patients With Severe Sepsis

Background—Increased plasma levels of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) have been identified as predictors of cardiac dysfunction and prognosis in congestive heart failure and ischemic heart disease. In severe sepsis patients, however, no information is available yet about the prognostic value of natriuretic peptides. Therefore, the aim of the present study was to determine the role of the N-terminal prohormone forms of ANP (NT-proANP) and BNP (NT-proBNP) in the context of outcome of septic patients. Furthermore, the effect of treatment with recombinant human activated protein C [drotrecogin alfa (activated)] on plasma levels of natriuretic peptides in severe sepsis was evaluated. Methods and Results—Fifty-seven patients with severe sepsis were included. Levels of NT-proANP and NT-proBNP were measured on the second day of sepsis by ELISA. Septic patients with NT-proBNP levels >1400 pmol/L were 3.9 times more likely (relative risk [RR], 3.9; 95% CI, 1.6 to 9.7) to die from sepsis than patients with lower NT-proBNP values (P<0.01). NT-proANP levels, however, were not predictive of survival in our patient population. A highly significant correlation was found between troponin I levels and plasma concentrations of NT-proBNP in septic patients (r=0.68, P<0.0001). In addition, troponin I significantly accounted for the variation in NT-proBNP levels (P<0.0001), suggesting an important role for NT-proBNP in the context of cardiac injury and dysfunction in septic patients. Twenty-three septic patients who received treatment with drotrecogin alfa (activated) presented with significantly lower concentrations of NT-proANP, NT-proBNP, and troponin I compared with patients not receiving drotrecogin alfa (activated). Conclusions—NT-proBNP may serve as useful laboratory marker to predict survival in patients presenting with severe sepsis.

Interleukin-1 beta promotes matrix metalloproteinase expression and cell proliferation in calcific aortic valve stenosis

Calcific aortic valve stenosis (AS), the main heart valve disease in the elderly, is characterized by extensive remodeling of the extracellular matrix. Matrix metalloproteinases (MMPs) are upregulated in calcific AS and might modulate matrix remodeling. The regulatory mechanisms are unclear. As recent studies have suggested that calcific AS might result from an inflammatory process involving leukocyte invasion and activation, the present study aimed to elucidate the role of the pro-inflammatory cytokine interleukin (IL)-1 beta on MMP expression and cell proliferation in human aortic valves. Immunohistochemistry for leukocytes, IL-1 beta and MMP-1 was performed on aortic valves with (n=6) and without (n=6) calcification obtained at valve replacement or autopsy. Stenotic valves showed marked leukocyte infiltration and associated expression of IL-1 beta and MMP-1. In control valves only scattered leukocytes, low staining for MMP-1 and no staining for IL-1 beta were present. Double-label immunostaining localized IL-1 beta expression mainly to leukocytes and MMP-1 expression to myofibroblasts. Stimulation of cultured human aortic valve myofibroblasts with IL-1 beta lead to a time-dependently increased expression of MMP-1 and MMP-2 by Western blotting and zymography, whereas MMP-9 remained unchanged. Cell proliferation was increased by IL-1 beta as determined by bromodesoxyuridine incorporation. Thus, IL-1 beta may regulate remodeling of the extracellular matrix in calcific AS.

Percutaneous coronary excimer laser angioplasty in patients with stable and unstable angina pectoris. Acute results and incidence of restenosis during 6-month follow-up.

A clinical study was conducted to evaluate the efficacy and safety of percutaneous coronary excimer laser angioplasty in 60 patients with coronary artery disease. Forty-nine patients had stable exertional angina, and 11 patients had unstable angina despite medical therapy. A novel 1.4-mm diameter catheter with 20 quartz fibers of 100-microns diameter each arranged concentrically around a central lumen suitable for a 0.014-in. flexible guide wire was coupled to an excimer laser. A commercial excimer laser emitting energy at a wavelength of 308 nm with a pulse duration of 60 nsec was used. The laser was operated at 20 Hz. Mean energy transmission was 30 +/- 5 mJ/mm2. In five of the 60 patients, laser angioplasty was not attempted. In 23 patients with laser ablation alone, percent stenosis decreased from 76 +/- 14% before to 27 +/- 17% after ablation and was 34 +/- 15% at the early follow-up angiogram. In 32 patients, additional balloon angioplasty was performed because of vessel closure after laser ablation in 11 and an insufficient qualitative result in 21 patients. Of the 11 patients with unstable angina, one patient died due to vessel closure 3 hours after intervention, and two patients developed a myocardial infarction. In 22 of 47 patients with late follow-up angiography, restenosis within the 6-month follow-up period occurred. Rate of restenosis was higher in patients treated with laser ablation and balloon angioplasty (16 of 28) than in patients treated with laser ablation alone (six of 19). These results suggest that coronary excimer laser angioplasty for ablation of obstructive lesions is feasible and safe in patients with stable angina. However, development of new catheter systems is necessary for an improved success rate.

Smooth muscle cell proliferation and restenosis after stand alone coronary excimer laser angioplasty

It has been shown that coronary excimer laser angioplasty can remove atherosclerotic intracoronary tissue. Stand alone coronary excimer laser angioplasty was successfully performed in a 53 year old white man with 90% stenosis of the left anterior descending coronary artery and exertional angina (Canadian Cardiovascular Society class III). The lesion was reduced to a 30% residual stenosis with use of a 1.2 mm and subsequently a 1.8 mm diameter laser catheter. Early follow-up angiography 24 h later revealed persistent patency and unchanged lesion diameter of the target vessel. The patient was free of symptoms during the 2 month follow-up period, but died suddenly while playing in a tennis tournament 63 days after the procedure. Postmortem histologic examination revealed 80% restenosis at the lesion site without plaque disruption or thrombosis. Specific staining of the histologic specimen for smooth muscle cells using alpha-actin revealed significant smooth muscle cell proliferation at the site of coronary excimer laser angioplasty. However, most of the vessel narrowing appeared to be due to underlying fibrotic plaque as a result of insufficient tissue ablation. This was probably related to the size of the currently available catheters, which are too small to create a large channel.

Drug eluting stents-potential applications for peripheral arterial occlusive disease

Many different approaches have been evaluated to prevent restenosis in stents after vascular implantation. Currently, drug-eluting stents are extremely promising in suppressing neointimal hyperplasia. Various animal studies and randomized trials in humans have shown excellent results in terms of safety and efficacy during intermediate-term follow-up. This article will give an overview of experimental and clinical data of the different agents in published and ongoing trials.

In vivo intravascular electric impedance spectroscopy using a new catheter with integrated microelectrodes.

Abstract Interventional techniques are necessary, which allow the characterization of intravascular pathological processes. Electric impedance spectroscopy (EIS) can provide cellular information of biological tissue. We tested the feasibility of intravascular EIS by using a new impedance catheter system with integrated microelectrodes in an experimental animal model. Eighteen stents were implanted into the iliac arteries of female New Zealand White rabbits (n = 11) to induce intimal proliferation. After 14, 28 and 56 days the electric impedance was measured inside and outside of the stented arterial segments by using a balloon catheter with four integrated microelectrodes. The impedance was recorded at a frequency ranging from 1 Hz to 1 MHz. After the measurements, the stents were explanted and histomorphometry was performed. The impedance inside and outside the stent was analysed and compared with the histomorphometric data.Fourteen (n = 6), 28 (n = 5) and 56 (n = 6) days after stent implantation the difference of the electrical impedance between the native and the stented iliac artery segment increased from –924 ± 715 Ohm to 3689 ± 1385 Ohm (14 days vs. 28 days; p < 0.05) and 8637 ± 2881 Ohm (14 days vs. 56 days; p < 0.05), respectively. The increase of the electrical impedance corresponded to an increased neointimal proliferation in the stented arterial segment of 3.6% ± 0.7% after 14 days, 8.4% ± 4.8% after 28 days (14 days vs. 28 days; p < 0.05) and 10.0% ± 4.1% after 56 days (14 days vs. 56 days; p < 0.01).Intravascular EIS can be performed by a balloon catheter with integrated microelectrodes and allows the detection of neointimal proliferation after stent implantation.

Intravascular electric impedance spectroscopy of atherosclerotic lesions using a new impedance catheter system

Newer techniques are required to identify atherosclerotic lesions that are prone to rupture. Electric impedance spectroscopy (EIS) can characterize biological tissues by measuring the electrical impedance over a frequency range. We tested a newly designed intravascular impedance catheter (IC) by measuring the impedance of different stages of atherosclerosis induced in an animal rabbit model. Six female New Zealand White rabbits were fed for 17 weeks with a 5% cholesterol–enriched diet to induce early forms of atherosclerotic plaques. All aortas were prepared from the aortic arch to the renal arteries and segments of 5–10 mm were marked by ink spots. A balloon catheter system with an integrated polyimide–based microelectrode structure was introduced into the aorta and the impedance was measured at each spot by using an impedance analyzer. The impedance was measured at frequencies of 1 kHz and 10 kHz and compared with the corresponding histomorphometric data of each aortic segment.Forty–four aortic segments without plaques and 48 segments with evolving atherosclerotic lesions could be exactly matched by the histomorphometric analysis. In normal aortic segments (P0) the change of the magnitude of impedance at 1 kHz and at 10 kHz (|Z|1 kHz – |Z|10 kHz, = ICF) was 208.5 ± 357.6 Ω. In the area of aortic segments with a plaque smaller than that of the aortic wall diameter (PI), the ICF was 137.7 ± 192.8 Ω. (P 0 vs. P I; p = 0.52), whereas in aortic segments with plaque formations larger than the aortic wall (PII) the ICF was significantly lower –22.2 ± 259.9 Ω. (P0 vs. PII; p = 0.002). Intravascular EIS could be successfully performed by using a newly designed microelectrode integrated onto a conventional coronary balloon catheter. In this experimental animal model atherosclerotic aortic lesions showed significantly higher ICF in comparison to the normal aortic tissue.

Time-dependent changes in the plasma concentration of matrix metalloproteinase 9 after acute myocardial infarction

Matrix metalloproteinase (MMP)-2 and MMP-9 are believed to play a pathophysiologic role in acute myocardial infarction (MI). The time course of their plasma concentrations in correlation with the extent of myocardial damage is unclear. In a prospective study, 20 patients with proven acute MI underwent successful reperfusion within 6 h after the onset of symptoms. The patients were divided into two groups according to the size of their MI, i.e. large or moderate MI. Plasma concentrations of MMP-2, MMP-9 and tissue inhibitor of metalloproteinase (TIMP)-1 were determined on admission, and after 24 h, 48 h, 1 week, 4 weeks, 3 months and 6 months. MMP-2 levels remained unchanged over time in both groups. The plasma concentration of MMP-9 was elevated on admission in patients with large MI versus moderate MI (195 ± 190 versus 78 ± 63 ng/ml, p < 0.01) as determined by left ventriculography, and returned to baseline (18 ± 16 ng/ml) by 1 week after MI. TIMP-1 levels rose slowly in patients with large MI and returned to baseline at 6 months. The ratio of MMP-9 to TIMP-1 was significantly increased on admission in both groups and returned to baseline at 48 h. These data suggest that MMP-9 might play a pathophysiologic role during the early phase of acute MI.

Local drug delivery: Impact of pressure, substance characteristics, and stenting on drug transfer into the arterial wall

Injection parameters for local drug delivery are frequently determined by studies with marker substances. However, the pharmacologic properties of the actual drug may influence delivery efficiency and lead to different results. Aim of this study was to assess the delivery capacities of two device‐drug combinations in order to verify this approach for further in vivo studies. Tritiated (3H) preparations (5 ml) of the hydrophylic low‐molecular‐weight heparin reviparin and the lipophilic taxane paclitaxel were injected into the left anterior descending artery of freshly explanted porcine hearts with the Infusasleeve II catheter system. A balloon support pressure of 6 atm and infusion pressures of 40, 60, 80, or 100 psi were used. In three additional groups, reviparin was injected following stent implantation and paclitaxel was injected prior to or following stent implantation. Arteries along with surrounding myocardium were harvested. The artery was carefully dissected, and artery and myocardium were separately homogenized, and activity was measured. Of the totally delivered activity, 0.09% ± 0.03% (40 psi) to 0.17% ± 0.13% (100 psi) of reviparin and 2.03% ± 0.67% (60 psi) to 2.68% ± 1.57% (100 psi) of paclitaxel were found in the vessel wall. The results for different injection pressures were not significantly different for either drug. The percentage activity delivered to the vessel wall was substantially larger in the paclitaxel group as compared to reviparin delivery (P < 0.01 at 60, 80, and 100 psi). The mean concentration of reviparin in the artery was 20 to 33 times higher than in the myocardium. For paclitaxel the factors were 110 to 243. Stent implantation prior to or following local delivery did not result in a different delivery efficiency. The results demonstrate that the characteristics of the delivered drug contribute largely to the delivery efficiency. Using identical injection parameters, drug concentrations in the arterial wall were significantly higher for the lipophilic paclitaxel as compared to the hydrophilic reviparin. Stenting of the artery did not influence delivery efficiency. Cathet. Cardiovasc. Intervent. 47:102–106, 1999.

Expression and activity of matrix metalloproteinase-2 in calcific aortic stenosis.

Die degenerativ- kalzifizierende Aortenstenose ist die häufigste Herzklappenerkrankung und Hauptursache eines Herzklappenersatzes im fortgeschrittenen Alter. Sie führt zu massiver Verkalkung sowie zu einem extensivem Umbau der extrazellulären Matrix; es wird vermutet, dass Matrix-Metalloproteinasen (MMPs) hierbei eine pathogenetische Rolle spielen. Wir untersuchten daher die Expression der Gelatinase MMP-2 und MMP-9 sowie ihres endogenen Inhibitors „Tissue Inhibitor of Metalloproteinase-2“ (TIMP-2) sowie die gelatinolytische Aktivität in 24 stenotischen und 8 normalen Aortenklappen. In der immunhistochemischen Färbung zeigten die stenotischen Klappen eine signifikant höhere Färbeintensität für MMP-2 und TIMP-2 im Vergleich zu den Kontrollklappen. Eine geringe Färbung von MMP-9 war ausschließlich bei stenotischen Klappen nachweisbar. In der in situ Zymographie wiesen normale Klappen eine minimale basale gelatinolytische Aktivität auf, die durch Zugabe des MMP-Aktivators p-Aminophenymercuroazetat (APMA) signifikant gesteigert werden konnte. In stenotischen Klappen war hingegen eine deutlich vermehrte Enzymaktivität nachweisbar, die durch Zugabe von APMA nicht mehr signifikant gesteigert werden konnte. MMP-2 und TIMP-2 zeigen somit eine differentielle Expression bei kalzifizierender Aortenstenose. MMP-2 liegt in normalen Klappen vorwiegend als inaktives Proenzym vor, in stenotischen Klappen hingegen in der aktivierten Form. Diese Ergebnisse sprechen für einen durch MMP-2 vermittelten Umbau der extrazellulären Matrix bei kalzifizierender Aortenstenose. Calcific aortic stenosis is the main heart valve disease in the elderly, leading to massive focal calcification and thickening of the valve cusps. Matrix metalloproteinases (MMPs) are thought to contribute to this process. Therefore, the study assessed the expression of the gelatinases MMP-2 and MMP-9 and the endogenous tissue inhibitor of metalloproteinase (TIMP)-2 as well as the gelatinolytic activity in normal and stenotic valves. Human tricuspid aortic valves with and without calcific aortic stenosis were studied by immunohistochemistry for MMP-2, MMP-9 and TIMP-2. The gelatinolytic activity in native valve sections was assessed by gelatin in situ zymography with or without addition of the MMP activator p-aminophenymercuric acetate (APMA). Staining intensities for MMP-2 and TIMP-2 were elevated in stenotic valves as compared to controls. Minor staining of MMP-9 was present exclusively in stenotic valves. The morphologic distribution of gelatinolytic activity was comparable to the staining pattern of MMP-2, and since MMP-9 immunostaining demonstrated only a low number of positive cells, the observed gelatinolytic activity is likely due to MMP-2. Gelatinolytic activity was low in normal valves but significantly increased by the MMP activator APMA. In contrast, stenotic valves showed a strong basal gelatinolytic activity that could not be significantly enhanced by APMA suggesting that MMP-2 is present as a latent pro-enzyme in normal valves and activated in stenotic valves. Thus, MMP-2 might be involved in the matrix remodeling during calcific aortic stenosis.